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Actin and hnRNP U cooperate for productive transcription by RNA polymerase II

Nature Structural & Molecular Biology volume 12, pages 238–244 (2005)Cite this article

Abstract

To determine the role of actin–ribonucleoprotein complexes in transcription, we set out to identify novel actin-binding proteins associated with RNA polymerase II (Pol II). Using affinity chromatography on fractionated HeLa cells, we found that hnRNP U binds actin through a short amino acid sequence in its C-terminal domain. Post-transcriptional gene silencing of hnRNP U and nuclear microinjections of a short peptide encompassing the hnRNP U actin-binding sequence inhibited BrUTP incorporation in vivo. In living cells, we found that both actin and hnRNP U are associated with the phosphorylated C-terminal domain of Pol II, and antibodies to actin and hnRNP U blocked Pol II–mediated transcription. Taken together, our results indicate that a general actin-based mechanism is implicated in the transcription of most Pol II genes. Actin in complex with hnRNP U may carry out its regulatory role during the initial phases of transcription activation.

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Figure 1: Actin binds the C-terminal domain of hnRNP U.
Figure 2: The actin-hnRNP U interaction takes place through a conserved motif located in the C-terminal domain of hnRNP U.
Figure 3: Post-transcriptional gene silencing of hnRNP U produces a considerable decrease in BrUTP incorporation.
Figure 4: In vivo disruption of the actin-hnRNP U interaction inhibits transcription.
Figure 5: Actin and hnRNP U are associated with the Pol II phosphoCTD in living cells.

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Acknowledgements

We thank B. Daneholt, N. Visa and D. Nashchekin for critically reading this manuscript. This work was supported by grants from the Swedish Research Council, the Swedish Cancer Society (project 4159), the Lars Hiertas Minne Foundation and the Jeansson Foundation to P.P. A.K. is a recipient of a doctoral fellowship from the Swedish Institute.

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Author notes

  1. Alexander Kukalev and Ylva Nord: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cell and Molecular Biology, Medical Nobel Institute, Karolinska Institute, Stockholm, S-171 77, Sweden

    Alexander Kukalev, Ylva Nord & Piergiorgio Percipalle

  2. Departments of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, S-171 77, Sweden

    Carina Palmberg & Tomas Bergman

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  1. Alexander Kukalev
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Correspondence to Piergiorgio Percipalle.

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Supplementary information

Supplementary Fig. 1

Post-transcriptional gene silencing of hnRNP U by RNAi. (PDF 223 kb)

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Kukalev, A., Nord, Y., Palmberg, C. et al. Actin and hnRNP U cooperate for productive transcription by RNA polymerase II. Nat Struct Mol Biol 12, 238–244 (2005). https://doi.org/10.1038/nsmb904

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